protein interaction by pull down

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zepedrof
zepedrof's picture
protein interaction by pull down

Hi all,

I have to test protein interaction by the pull down method using one protein fuised do his tag and another protein fused to GST tag. I have successfully induced expression of both proteins but one is soluble while the other is unsoluble.

can I do pull down of this unsoluble protein ?  or do i need to refold the protein first ?

thanks,

JP

Satinder
Satinder's picture
refolding would be a better

refolding would be a better choice for allowing the solubility of the protein. Other wise try a higher concentration of detergents for the pull down assay. You can also incorporate urea in yr initial solubilization step and after solubilization of the same reduce its concentrtion by addition of a buffer suitable  for pull dowwn.